Abstract
Plasmid-mediated quinolone resistance mechanisms have become increasingly prevalent among Enterobacteriaceae strains since the 1990s. Among these mechanisms, AAC(6’)-Ib-cr is the most difficult to detect. Different detection methods have been developed, but they require expensive procedures such as Sanger sequencing, pyrosequencing, polymerase chain reaction (PCR) restriction, or the time-consuming phenotypic method of Wachino. In this study, we describe a simple matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) method which can be easily implemented in clinical laboratories that use the MALDI-TOF technique for bacterial identification. We tested 113 strains of Enterobacteriaceae, of which 64 harbored the aac(6’)-Ib-cr gene. We compared two MALDI-TOF strategies, which differed by their norfloxacin concentration (0.03 vs. 0.5 g/L), and the method of Wachino with the PCR and sequencing strategy used as the reference. The MALDI-TOF strategy, performed with 0.03 g/L norfloxacin, and the method of Wachino yielded the same high performances (Se = 98 %, Sp = 100 %), but the turnaround time of the MALDI-TOF strategy was faster (<5 h), simpler, and inexpensive (<1 Euro). Our study shows that the MALDI-TOF strategy has the potential to become a major method for the detection of many different enzymatic resistance mechanisms.
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Acknowledgments
We thank Laurent Guillouard and Alexis Pontvianne for their technical assistance. This work was presented in part at the 31ème Réunion Interdisciplinaire de Chimiothérapie Anti-Infectieuse (RICAI), Paris, France, December 2011.
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This work was supported by a grant from the French Ministry of Health via the Institut de Veille Sanitaire and by a grant from the Centre Hospitalier Régional Universitaire de Clermont-Ferrand, France
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All authors report no conflicts of interest in relation to this article.
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Pardo, CA., Tan, R.N., Hennequin, C. et al. Rapid detection of AAC(6’)-Ib-cr production using a MALDI-TOF MS strategy. Eur J Clin Microbiol Infect Dis 35, 2047–2051 (2016). https://doi.org/10.1007/s10096-016-2762-1
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DOI: https://doi.org/10.1007/s10096-016-2762-1